Fetal alcohol syndrome is recognized as the leading known cause of mental retardation in the Western World. Fetal alcohol syndrome is characterized by abnormal brain development, hippocampus, and cerebral cortex being the most affected areas in the developing brain. There are a number of deficits which occur such as prenatal and postnatal deficiencies, microcephaly, and craniofacial abnormalities. While we do not know the underlying mechanism of enhanced cell death in the ethanol exposed fetal brain, there is increasing evidence that ethanol induces an increase in apoptotic cell death. It is hypothesized that ex utero ethanol exposure initiates a cascade of events at the mitochondrial level in the developing brain, specifically by direct effects on astrocytes and neurons, inducing oxidative damage and ultimately apoptotic cell death. These studies will utilize three models including the "pup in a cup" model of the brain growth spurt, and cultures of cortical neurons and astrocytes isolated from fetal and neonatal pups. The first specific aim is to determine the effects of ethanol exposure on the generation and location of reactive oxygen species in neonatal rat cortical astrocytes and primary cortical neurons. This will be accomplished through the use of cell viability studies, generation of ROS in intact cells and mitochondria. The second specific aim is designed to determine how ethanol induces apoptotic cell death in the developing neonatal rat brain using specific apoptotic measures. The apoptotic measures chosen are mitochondrial permeability transition, cytochrome c and AIF release, caspase-c activity, and DNA fragmentation.